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Article

Modulation of Gut–Liver Axis by ASD-Associated Microbiota and Synbiotic Intervention in a Pseudo-Germ-Free Mouse Model

by
Kristina Smajda Rodakova
1,†,
Sona Gancarcikova
2,†,
Vlasta Demeckova
3,
Stanislav Lauko
2,
Maria Rynikova
3,
Zuzana Andrejcakova
1,
Daniela Spisakova
2,
Michal Fusek
4,
Radoslava Vlckova
1,
Viola Strompfova
5,
Aleksandra Tomova
6,
Barbara Raskova
6 and
Drahomira Sopkova
1,*
1
Department of Biology and Physiology, University of Veterinary Medicine and Pharmacy in Kosice, 041 81 Kosice, Slovakia
2
Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Kosice, 041 81 Kosice, Slovakia
3
Department of Animal Physiology, Institute of Biology and Ecology, Faculty of Science, Pavol Jozef Safarik University in Kosice, 040 01 Kosice, Slovakia
4
Department of Mathematics, Faculty of Electrical Engineering and Communication, Brno University of Technology, 616 00 Brno, Czech Republic
5
Centre of Biosciences of the Slovak Academy of Sciences, Institute of Animal Physiology, Soltesovej 4-6, 040 01 Kosice, Slovakia
6
Institute of Physiology, Faculty of Medicine, Comenius University in Bratislava, 814 99 Bratislava, Slovakia
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Appl. Sci. 2026, 16(11), 5529; https://doi.org/10.3390/app16115529
Submission received: 17 April 2026 / Revised: 17 May 2026 / Accepted: 19 May 2026 / Published: 2 June 2026
(This article belongs to the Section Applied Microbiology)
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Abstract

Research on autism spectrum disorders (ASDs) has so far focused primarily on the gut–brain axis, whereas the role of the gut–liver axis remains insufficiently explored. The aim of this study was to evaluate whether microbiota derived from girls with ASD induces dysbiotic changes in gut microbiota composition and leads to alterations in gut–liver axis processes in pseudo-germ-free (PGF) BALB/c mice. We also examined whether these processes could be modulated by altering the gut microbiota using the probiotic strains Lactiplantibacillus plantarum CCM 7512 and Limosilactobacillus reuteri CCM 8617 in combination with ground flaxseed (Linum usitatissimum L.) as a source of omega-3 polyunsaturated fatty acids and fermentable fiber. Colonization with fecal microbiota derived from girls with ASD resulted in dysbiotic changes in the composition of the cecal microbiota, characterized by an increased relative abundance of Escherichia–Shigella, Fusobacterium, Alistipes, and the Ruminococcus gnavus group. These changes were accompanied by impaired intestinal mucosal integrity, altered metabolomic pathways related mainly to aromatic amino acids and lipid metabolism, increased hepatic immunoreactivity of iNOS and COX-2, and elevated activity of the liver-specific LDH-5 isoenzyme. These results suggest that synbiotic intervention contributed to remodeling of the cecal microbiota composition, restoration of intestinal epithelial integrity, and modulation of metabolomic pathways, which was reflected by reduced immunoreactivity of iNOS and COX-2 in liver tissue and decreased activity of the LDH-5 isoenzyme. These findings support the role of microbiota-mediated metabolic processes in communication between the gut and the liver within the gut–liver axis.
Keywords: autism spectrum disorder; gut microbiota; gut–liver axis; fecal microbiota transplantation; pseudo-germ-free mice; metabolomics; probiotics; prebiotics autism spectrum disorder; gut microbiota; gut–liver axis; fecal microbiota transplantation; pseudo-germ-free mice; metabolomics; probiotics; prebiotics

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MDPI and ACS Style

Rodakova, K.S.; Gancarcikova, S.; Demeckova, V.; Lauko, S.; Rynikova, M.; Andrejcakova, Z.; Spisakova, D.; Fusek, M.; Vlckova, R.; Strompfova, V.; et al. Modulation of Gut–Liver Axis by ASD-Associated Microbiota and Synbiotic Intervention in a Pseudo-Germ-Free Mouse Model. Appl. Sci. 2026, 16, 5529. https://doi.org/10.3390/app16115529

AMA Style

Rodakova KS, Gancarcikova S, Demeckova V, Lauko S, Rynikova M, Andrejcakova Z, Spisakova D, Fusek M, Vlckova R, Strompfova V, et al. Modulation of Gut–Liver Axis by ASD-Associated Microbiota and Synbiotic Intervention in a Pseudo-Germ-Free Mouse Model. Applied Sciences. 2026; 16(11):5529. https://doi.org/10.3390/app16115529

Chicago/Turabian Style

Rodakova, Kristina Smajda, Sona Gancarcikova, Vlasta Demeckova, Stanislav Lauko, Maria Rynikova, Zuzana Andrejcakova, Daniela Spisakova, Michal Fusek, Radoslava Vlckova, Viola Strompfova, and et al. 2026. "Modulation of Gut–Liver Axis by ASD-Associated Microbiota and Synbiotic Intervention in a Pseudo-Germ-Free Mouse Model" Applied Sciences 16, no. 11: 5529. https://doi.org/10.3390/app16115529

APA Style

Rodakova, K. S., Gancarcikova, S., Demeckova, V., Lauko, S., Rynikova, M., Andrejcakova, Z., Spisakova, D., Fusek, M., Vlckova, R., Strompfova, V., Tomova, A., Raskova, B., & Sopkova, D. (2026). Modulation of Gut–Liver Axis by ASD-Associated Microbiota and Synbiotic Intervention in a Pseudo-Germ-Free Mouse Model. Applied Sciences, 16(11), 5529. https://doi.org/10.3390/app16115529

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